4.2 Article

An improved method for the cost-effective expression and purification of large quantities of KcsA

期刊

PROTEIN EXPRESSION AND PURIFICATION
卷 127, 期 -, 页码 53-60

出版社

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2016.07.002

关键词

Ion channels; KcsA; Biochemistry; Over-expression; Detergents; Solubilization

资金

  1. CMPR-TTUHSC seeding grant
  2. American Heart Association [11SDG5440003]
  3. National Institute of Health [1RO1GM097159-01A1]
  4. Welch Foundation [BI-1757]

向作者/读者索取更多资源

KcsA, the bacterial K+ channel from Streptomyces lividans, is the prototypical model system to study the functional and structural correlations of the pore domain of eukaryotic voltage-gated K+ channels (K-v channels). It contains all the molecular elements responsible for ion conduction, activation, deactivation and inactivation gating [1]. KcsA's structural simplicity makes it highly amenable for structural studies. Therefore, it is methodological advantageous to produce large amounts of functional and properly folded KcsA in a cost-effective manner. In the present study, we show an optimized protocol for the over expression and purification of large amounts of high-quality, fully functional and crystallizable KcsA using inexpensive detergents, which significantly lowered the cost of the purification process. (C) 2016 Elsevier Inc. All rights reserved.

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